Patent AU-2021101572-A4 (AU)
Title: A method for producing surface modified silk fibroin nanoparticles for improved delivery of doxorubicin
Inventor: PANDEY VIKAS
IPC: A61K9/51
Filed: 2021-03-27
Granted: 2021-05-27
Abstract: The present disclosure relates to a method for producing surface modified silk fibroin 
nanoparticles for improved delivery of doxorubicin. Surface modified silk fibroin 
nanoparticles bearing anti-cancer agent doxorubicin (DOX) are fabricated by desolvation 
method and coated with Tween-80 as surface modifier. The prepared nanoparticles are 
characterized for various physicochemical parameters, like particle size, surface charge, 
surface morphology by scanning electron microscope (SEM) and transmission electron 
microscopy (TEM), and in vitro drug release along with in vitro cell cytotoxicity, flow 
cytometry and cellular uptake studies by flouro cytometry on glioblastoma cell lines.  
Entrapment efficiency for the silk fibroin nanoparticles are found to be >85% for coated and 
uncoated nanoparticles. Nanoparticles with average diameter less than 150nm having 
negative charge are found to show no toxicity of its own. The prion flammatory response of 
nanoparticles is observed by determining the cytokines level, such as TNF-a and IL.  
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cutingorchoppingtombyxmorisilkwormcocoonintosmallpiecesandboilingthesmallpiecesofcocon(5g)within0.02M 
solutionofsodium carbonate (NaC3)for60 min subsequentlyfollowed by whingseveraltimesith distilled cold ater 102 
squeezingwashedsilkfibrtoremoveexcesswaterandthereafterspreadingoutoncleanaluinum filpieefollwedbyairdrying 
dissolvingthedriedsilkfibrwinlithiumbromide[93MLBriat60degreeCelsusfor4 htoobtainambercdugrandtransparentwhich 106 
isfurtherdialyzedtoremvethelithiumbromide,whereinthedriedsilkfi resand ibristakeninthe1:4ratio(Ugdriedslkfibrto 
4ml9.3M 
preparingsilkfibroin nanopartIcles byd isolation or nanopredpitatIon method and addingsilkfibroinsoluton from Bombyx moi 108 
drop-wise(20pl/drop~toorgancsolventaetne,maintaining»75%v/vacetonevolume 
precipitatingaddedsilkfibroinsolutdonandthencentrifugingat4,40lxg for2htoconvertintnapelletfollowedbyseparationof A/ 
supernatantandthereafterthepelletformedduetoprecipitationofsilkfibroininaetoneandcentrifugationisthenre-susperidedin 110 
double distilledwater 
sonicatingthesuspensionat30%amplitudefor30secoodsbyaprobe soato 112 
adding100pgDOXsolutiontonanopartidesdispelsion(50pgsilkfibminnanoparbcleslwithcontinuousstirringof20revolutionsper 114 
minute[rpm||atroomtemperaturefor12hindarkconditionfordrugloadingtosilkfibroinnanopartdesandcentrifugatnat9000 
rpm for20 min toobtain DCX beangSFN (SFN@D ) 
suspending lyphilizedpowderofpreparedSFN @DOnanopartidesinPBS(pH7.4(andcoatingwithTween-80toformTSPFN@D 116 
andadding0.S% v/vTween-80relativetototalsuspensionvolumeofnanoparticles 
incubatingmixtureofSFN@D midth0.5%v/vTween-20isfor3Ominat37degreeCeliusandperforminglyophiliationofthe 
surface modified nanoparticlesTSFN@DXtoyield nanopartidesin powdered form 
Figure1 
Add silk fibrAinsolution f 
0.02 M N aCOu dropwIse (20pt/drop) 
Cutting 93 M LBr 
Slik broin 
mby mrnfi siCk worr ut pices of Brbyx 
solution 
yxor ssk worm cocoons 
cocoons Acetore (175% vn) 
Silik Fib-.1-0I 
Ne^norrtklos (SFN) 
Add DOX a tlui S 
to SFN 
Add ~Centnlfugatlo > 
dd0.5%v ween-o (48400 X for 20 min) 
Lyophillzed powder of 
SPN@DOX 
DOX barin SFNDOX bearing SFN 
(TS )(SFN DOX) Stlring at 20 rpm for 12 h 
Figure 2